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Formulation of microbial cocktails for BTEX biodegradation

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Abstract

BTEX biodegradation by a mixed community of micro-organisms offers a promising approach in terms of cost-effectiveness and elimination of secondary pollution. Two bacterial strains, Pseudomonas putida F1 and Pseudomonas stutzeri OX1 were chosen to formulate synthetic consortia based on their ability to biodegrade the mono-aromatic compounds. Benzene and toluene supported the growth of both the strains; while ethyl benzene and o-xylene were only utilized as growth substrates by P. putida F1 and P. stutzeri OX1, respectively. In a mixed substrate system, P. putida F1 exhibited incomplete removal of o-xylene while P. stutzeri OX1 displayed cometabolic removal of ethyl benzene with dark coloration of the growth medium. The biodegradation potential of the two Pseudomonas species complemented each other and offered opportunities to explore their performance as a co-culture for enhanced BTEX biodegradation. Several microbial formulations were concocted and their BTEX biodegradation characteristics were evaluated. Mixed culture biodegradation ascertained the advantages of the co-culture over the individual Pseudomonas species. This study also emphasized the significance of inoculum density and species proportion while concocting preselected micro-organisms for enhanced BTEX biodegradation.

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References

  • Abuhamed T, Bayraktar E, Mehmetoğlu T, Mehmetoğlu Ü (2004) Kinetics model for growth of Pseudomonas putida F1 during benzene, toluene and phenol degradation. Process Biochem 39:983–988

    Article  CAS  Google Scholar 

  • Arafa MA (2003) Biodegradation of some aromatic hydrocarbons (BTEXs) by a bacterial consortium isolated from polluted site in Saudi Arabia. Pak J Biol Sci 6:1482–1486

    Article  Google Scholar 

  • Attaway HH, Schmidt MG (2002) Tandem biodegradation of BTEX components by two Pseudomonas sp. Curr Microbiol 45:30–36

    Article  CAS  PubMed  Google Scholar 

  • Baggi G, Barbieri P, Galli E, Tollari S (1987) Isolation of a Pseudomonas stutzeri strain that degrades o-xylene. Appl Environ Microbiol 53:2129–2132

    CAS  PubMed Central  PubMed  Google Scholar 

  • Barbieri P, Arenghi FLG, Bertoni G, Bolognese F, Galli E (2001) Evolution of catabolic pathways and metabolic versatility in Pseudomonas stutzeri OX1. Antonie Van Leeuwenhoek 79:135–140

    Article  CAS  PubMed  Google Scholar 

  • Barbieri P, Solera D, Galli E, Vidal-Aroca F, Bertoni G (2007) Degradation of o-xylene by Pseudomonas stutzeri OX1 (Pseudomonas sp. OX1). In: Ramos J-L, Filloux A (eds) Pseudomonas A model system in biology, vol 2. Springer, New York, pp 89–105

    Google Scholar 

  • Bielefeldt AR, Stensel HD (1999) Modeling competitive inhibition effects during biodegradation of BTEX mixtures. Water Res 33:704–714

    Google Scholar 

  • Chakraborty R, Coates JD (2004) Anaerobic degradation of monoaromatic hydrocarbons. Appl Microbiol Biotechnol 64:437–446

    Article  CAS  PubMed  Google Scholar 

  • Chang M-K, Voice TC, Criddle CS (1993) Kinetics of competitive inhibition and cometabolism in the biodegradation of benzene, toluene and p-xylene by two Pseudomonas isolates. Biotechnol Bioeng 41:1057–1065

    Article  CAS  Google Scholar 

  • Deeb RA, Alvarez-Cohen L (1999) Temperature and substrate interactions during the aerobic biotransformation of BTEX mixtures by toluene-enriched consortia and Rhodococcus rhodochrous. Biotechnol Bioeng 62:526–536

    Article  CAS  PubMed  Google Scholar 

  • Díaz E (2004) Bacterial degradation of aromatic pollutants: a paradigm of metabolic versatility. Int Microbiol 7:173–180

    PubMed  Google Scholar 

  • Dou J, Liu X, Hu Z (2008) Anaerobic BTEX degradation in soil biaugmented with mixed consortia under nitrate reducing conditions. J Environ Sci 20:585–592

    Article  CAS  Google Scholar 

  • Duetz WA, Wind B, van Andel JG, Barnes MR, Williams PA, Rutgers M (1998) Biodegradation kinetics of toluene, m-xylene, p-xylene and their intermediates through the upper TOL pathway in Pseudomonas putida (PWWO). Microbiology 144:1669–1675

    Article  CAS  Google Scholar 

  • Fries MR, Zhou J, Chee-Sanford J, Tiedje JM (1994) Isolation, characterization, and distribution of denitrifying toluene degraders from a variety of habitats. Appl Environ Microbiol 60:2802–2810

    CAS  PubMed Central  PubMed  Google Scholar 

  • Gilbert ES, Walker AW, Keasling JD (2003) A constructed microbial consortium for biodegradation of the organophosphorus insecticide parathion. Appl Microbiol Biotechnol 61:77–80

    Article  CAS  PubMed  Google Scholar 

  • Jo MS, Rene ER, Kim SH, Park HS (2008) An analysis of synergistic and antagonistic behavior during BTEX removal in batch system using response surface methodology. J Hazard Mater 152:1276–1284

    Article  CAS  PubMed  Google Scholar 

  • Jung I-G, Park C-H (2004) Characteristics of Rhodococcus pyridinovorans PYJ-1 for the biodegradation of benzene, toluene, m-xylene (BTX), and their mixtures. J Biosci Bioeng 97:429–431

    Article  CAS  PubMed  Google Scholar 

  • Kahraman H, Geckil H (2005) Degradation of benzene, toluene and xylene by Pseudomonas aeruginosa engineered with the Vitreoscilla Hemoglobin gene. Eng Life Sci 5:363–368

    Article  CAS  Google Scholar 

  • Kim D-J, Choi J-W, Choi N-C, Mahendran B, Lee C-E (2005) Modeling of growth kinetics for Pseudomonas spp. during benzene degradation. Appl Microbiol Biotechnol 69:456–462

    Article  CAS  PubMed  Google Scholar 

  • Lee E, Cho KS (2009) Effect of substrate interaction on the degradation of methyl tert-butyl ether, benzene, toluene, ethylbenzene, and xylene by Rhodococcus sp. J Hazard Mater 167:669–674

    Article  CAS  PubMed  Google Scholar 

  • Lee S-K, Lee SB (2001) Isolation and characterization of a thermotolerant bacterium Ralstonia sp. strain PHS1 that degrades benzene, toluene, ethylbenzene, and o-xylene. Appl Microbiol Biotechnol 56:270–275

    Article  CAS  PubMed  Google Scholar 

  • Lee J-Y, Roh JR, Kim H-S (1994) Metabolic engineering of Pseudomonas putida for the simultaneous biodegradation of benzene, toluene, and p-xylene mixture. Biotechnol Bioeng 43:1146–1152

    Article  CAS  PubMed  Google Scholar 

  • Lee J-Y, Jung K-H, Choi SH, Kim H-S (1995) Combination of the tod and the tol pathways in redesigning a metabolic route of Pseudomonas putida for the mineralization of a benzene, toluene, and p-xylene mixture. Appl Environ Microbiol 61:2211–2217

    CAS  PubMed Central  PubMed  Google Scholar 

  • Littlejohns JV, Daugulis AJ (2008) Kinetics and interactions of BTEX compounds during degradation by a bacterial consortium. Process Biochem 43:1068–1076

    Article  CAS  Google Scholar 

  • Mathur AK, Majumder CB (2010) Kinetics modelling of the biodegradation of benzene, toluene and phenol as single substrate and mixed substrate by using Pseudomonas putida. Chem Biochem Eng Q 24:101–109

    CAS  Google Scholar 

  • Mazzeo DEC, Levy CE, de Angelis DDF, Marin-Morales MA (2010) BTEX biodegradation by bacteria from effluents of petroleum refinery. Sci Total Environ 408:4334–4340

    Article  CAS  PubMed  Google Scholar 

  • Muñoz R, Díaz LF, Bordel S, Villaverde S (2007) Inhibitory effects of catechol accumulation on benzene biodegradation in Pseudomonas putida F1 cultures. Chemosphere 68:244–252

    Article  PubMed  Google Scholar 

  • Oh Y-S, Bartha R (1997) Construction of a bacterial consortium for the biofiltration of benzene, toluene and xylene emissions. World J Microbiol Biotechnol 13:627–632

    Article  CAS  Google Scholar 

  • Oh Y-S, Choi SC (1997) Characterization of BTX-degrading bacteria and identification of substrate interactions during their degradation. J Microbiol 35:193–199

    CAS  Google Scholar 

  • Oh Y-S, Shareefdeen Z, Baltzis BC, Bartha R (1994) Interactions between benzene, toluene and p-xylene (BTX) during their biodegradation. Biotechnol Bioeng 44:533–538

    Article  CAS  PubMed  Google Scholar 

  • Otenio MH, Da Silva MTL, Marques MLO, Roseiro JC, Bidoia ED (2005) Benzene, toluene and xylene biodegradation by Pseudomonas putida CCMI 852. Braz J Microbiol 36:258–261

    Article  CAS  Google Scholar 

  • Prenafeta-Boldú FX, Vervoot J, Grotenhuis JTC, Van Groenestijn JW (2002) Substrate interactions during the biodegradation of benzene, toluene, ethylbenzene, and xylene (BTEX) hydrocarbons by the fungus Cladophialophora sp. strain T1. Appl Environ Microbiol 68:2660–2665

    Article  PubMed Central  PubMed  Google Scholar 

  • Reardon KF, Mosteller DC, Rogers JB (2000) Biodegradation kinetics of toluene and phenol as single and mixed substrates for Pseudomonas putida F1. Biotechnol Bioeng 69:385–400

    Article  CAS  PubMed  Google Scholar 

  • Rogers JB, Reardon KF (2000) Modelling substrate interactions during the biodegradation of mixtures of toluene and phenol by Burkholderia species JS150. Biotechnol Bioeng 70:428–435

    Article  CAS  PubMed  Google Scholar 

  • Tsao C-W, Song H-G, Bartha R (1998) Metabolism of benzene, toluene and xylene hydrocarbons in soil. Appl Environ Microbiol 64:4924–4929

    CAS  PubMed Central  PubMed  Google Scholar 

  • Yu H, Kim BJ, Rittmann BE (2001) The role of intermediates in biodegradation of benzene, toluene and p-xylene by Pseudomonas putida F1. Biodegradation 12:455–463

    Article  CAS  PubMed  Google Scholar 

  • Zylstra GJ (1994) Molecular analysis of aromatic hydrocarbon degradation. In: Garte SJ (ed) Molecular environmental biology. Lewis Publishers, Boca Raton

    Google Scholar 

  • Zylstra GJ, McCombie WR, Gibson DT, Finette BA (1988) Toluene degradation by Pseudomonas putida F1: genetic organization of the tod operon. Appl Environ Microbiol 54:1498–1503

    CAS  PubMed Central  PubMed  Google Scholar 

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Correspondence to Kai-Chee Loh.

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Nagarajan, K., Loh, KC. Formulation of microbial cocktails for BTEX biodegradation. Biodegradation 26, 51–63 (2015). https://doi.org/10.1007/s10532-014-9715-0

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  • DOI: https://doi.org/10.1007/s10532-014-9715-0

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